A multi-parametric flow cytometric assay to analyze DNA–protein interactions

Interactions between DNA and transcription factors (TFs) guide cellular function and development, yet the complexities of gene regulation are still far from being understood. Such understanding is limited by a paucity of techniques with which to probe DNA–protein interactions. We have devised magnetic protein immobilization on enhancer DNA (MagPIE), a simple, rapid, multi-parametric assay using flow cytometric immunofluorescence to reveal interactions among TFs, chromatin structure and DNA. In MagPIE, synthesized DNA is bound to magnetic beads, which are then incubated with nuclear lysate, permitting sequence-specific binding by TFs, histones and methylation by native lysate factors that can be optionally inhibited with small molecules. Lysate protein–DNA binding is monitored by flow cytometric immunofluorescence, which allows for accurate comparative measurement of TF-DNA affinity. Combinatorial fluorescent staining allows simultaneous analysis of sequence-specific TF-DNA interaction and chromatin modification. MagPIE provides a simple and robust method to analyze complex epigenetic interactions in vitro

The ventilation effect on stator convective heat transfer of an axial-flux permanent-magnet machine

This paper investigates the effect of the inlet configuration on cooling for an air-cooled axial-flux permanent-magnet (AFPM) machine. Temperature rises in the stator were measured and compared with results predicted using computational fluid dynamic (CFD) methods linked to a detailed machine loss characterization. It is found that an improved inlet design can significantly reduce the stator temperature rises. Comparison between the validated CFD model results and the values obtained from heat transfer correlations addresses the suitability of those correlations proposed specifically for AFPM machines

Dementia with Lewy bodies: Impact of co-pathologies and implications for clinical trial design

Dementia with Lewy bodies (DLB) is clinically defined by the presence of visual hallucinations, fluctuations, rapid eye movement (REM) sleep behavioral disorder, and parkinsonism. Neuropathologically, it is characterized by the presence of Lewy pathology. However, neuropathological studies have demonstrated the high prevalence of coexistent Alzheimer's disease, TAR DNA-binding protein 43 (TDP-43), and cerebrovascular pathologic cases. Due to their high prevalence and clinical impact on DLB individuals, clinical trials should account for these co-pathologies in their design and selection and the interpretation of biomarkers values and outcomes. Here we discuss the frequency of the different co-pathologies in DLB and their cross-sectional and longitudinal clinical impact. We then evaluate the utility and possible applications of disease-specific and disease-nonspecific biomarkers and how co-pathologies can impact these biomarkers. We propose a framework for integrating multi-modal biomarker fingerprints and step-wise selection and assessment of DLB individuals for clinical trials, monitoring target engagement, and interpreting outcomes in the setting of co-pathologies.Wellcome Clinical Research Career Development Fellowship. Grant Number: 205167/Z/16/Z; National Institute for Health Research (NIHR); National Institute of Neurological Disorders and Stroke. Grant Number: 1ZIANS003154; NIH. Grant Numbers: P30 AG62677, U01 NS100620, U19 AG 071754, U54 NS110435 NINDS. Grant Number: U01NA100610; National Institute on Aging. Grant Number: P30AG072959Peer reviewe

Packages of Care for Alcohol Use Disorders in Low- And Middle-Income Countries

In the fourth in a series of six articles on packages of care for mental disorders in low- and middle-income countries, Vivek Benegal and colleagues discuss the treatment of alcohol use disorders